荧光定量PCR快速检测食品中肠出血性大肠杆菌O104

    刘雅婷1，杨宇2，王旺2，吉尚志2，刘丽娟2，曹刚强1，王静2

    1.郑州大学生物工程系，河南 郑州 450001；2.中国检验检疫科学研究院

    摘要：目的 建立针对肠出血性大肠杆菌O104的荧光定量PCR快速检测方法。方法针对O104的wzx保守基因，设计特异性引物和探针，采用倍比梯度稀释法检测该体系的灵敏度，以另外34株肠道致病菌评价检测方法的特异性；建立了肠出血性大肠杆菌O104感染食品的检测模型以验证方法的适用性。结果建立了荧光定量PCR快速检测肠出血性大肠杆菌O104的方法。每次检测最低限为12.0 copies，特异性为100%。结论该法缩短了检测时间，并有良好的灵敏性和特异性，在疾病防控及食品卫生行业中具有应用前景。

    关键词：肠出血性大肠杆菌O104；荧光定量PCR；检测；食品

    中图分类号：R155.5，R378.2+1　文献标识码：B

    Rapid detection of enterohemorrhagicescherichia coli O104 in food  with Real-time PCR assay

    LIU Ya-ting*, YANG Yu，WANG Wang， JIShang-zhi， LlU Li-juan， CAO Gang-qiang, WANG Jing

    *Department of Bio-engineer, ZhengzhouUniversity, Zhengzhou， Henan 450001, China

    Abstract:   Objective  To develop a real-time PCR assay for rapid detection ofenterohemorrhagic Escherichia coli O104.  Methods  The specific primers and probes were designed to amplify the wzxgene of EHEC O104. And the sensitivity of system was detected byusing fold dilution method. In order to examine the specificity ofthe system, other 34 intestinal bacteria strains were assayed andmimic EHEC O104 infected food was detected for evaluation. Results   A highly sensitive and specific real-time PCRassay was established for detection of EHEC O104. The sensitivitywas 12 copies/test for EHEC O104.The specificity was 100%. Conclusion   The real-time PCR detection method isefficient to detect EHEC O104 with good sensitivity andspecificity. The method has good prospects of application indisease prevention and food safety area.

    Key words:   EnterohemorrhagicEscherichia coli(EHEC) O104; Real-time PCR； Detection; Food

    《中国国境卫生检疫杂志》